Purification and Amino Terminal Sequencing of Human Melanoma Nerve Growth Factor Receptor

Marano, Nadia; Dietzschold, Bernhard; Jj, Earley; Schatteman, Gina C.; Thompson, Sumner E.; Grob, Peter M.; Ah, Ross; Bothwell, Mark; Bf, Atkinson; Koprowski, Hilary

doi:10.1111/j.1471-4159.1987.tb13151.x

Cited by 84 publications

(31 citation statements)

References 27 publications

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“…The ICso values presented here and in previous crosslinking studies (Vissavajjhala & Ross, 1990) were obtained with high RED concentration and, therefore, represent upper limits to the true dissociation constants for RED and the neurotro- KO. This value still represents a maximal estimate of the dissociation constant but is in reasonable agreement with that for LANR (K, about 1 nM) measured on the cell surface (Sutter et al, 1979;Vale & Shooter, 1985;Woodruff & Neet, 1986;Rodriguez-Tebar et al, 1990, 1992Squinto et al, 1991;Ip et al, 1992), measured with intact solubilized receptor (ECSo about 30 nM) (Marano et al, 1987), or measured with naturally occurring, truncated LANR (ECSo = 15 nM) (Zupan et al, 1989). This agreement between dissociation constants for binding to the purified, soluble extracellular domain and the intact receptor suggests the apparent affinity is not significantly affected by components in the cell membrane or by the receptor's cytosolic domain.…”

Section: Discussionsupporting

confidence: 60%

Circular dichroism and crosslinking studies of the interaction between four neurotrophins and the extracellular domain of the low‐affinity neurotrophin receptor

Timm

Ross²,

Neet

1994

Protein Science

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Interactions between the purified recombinant receptor extracellular domain (RED) of the human low-affinity neurotrophin receptor (LANR) and recombinant human brain-derived neurotrophic factor, neurotrophin-3 (NT-3) and neurotrophin-4/5 have been studied by chemical crosslinking and circular dichroism. Conformational changes subsequent to binding have been shown by these procedures. First, relative affinities of the neurotrophins for RED were determined by binding competition assays in which radioiodinated nerve growth factor (NGF) from mouse submaxillary gland was crosslinked to RED in the presence of varying amounts of unlabeled neurotrophin competitors. RED bound each of the 3 recombinant human neurotrophins with affinities that were indistinguishable from authentic mouse NGF. These results are the first measurement of binding of the neurotrophin family to their common receptor using purified components. In order to study the effect of binding on the conformation of the proteins, CD measurements were made before and after mixing neurotrophins and RED, as had previously been done with NGF and RED (Timm DE, Vissavajjhala P, Ross AH, Neet KE, 1992, Protein Sci 1:1023-1031). Similar changes in CD spectra occurred upon combination of each of the neurotrophins and RED, with negative changes near 220-225 nm and positive changes near 190-200 nm; however, significant differences existed among the various neurotrophin-RED difference spectra. The NT-3/RED complex showed the largest spectral change and NGF the smallest. Thus, specific conformational changes in secondary structure of neurotrophin, RED, or both accompany the binding of each neurotrophin to the extracellular domain of the LANR.Keywords: circular dichroism; conformational change; low-affinity NGF receptor; nerve growth factor; neurotrophin; receptorThe neurotrophins are a family of target-derived trophic factors required for the development and survival of specific neuronal populations (Levi-Montalcini, 1987). This family of homologous proteins includes nerve growth factor, brain-derived neuro-

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Section: Discussionsupporting

confidence: 60%

Circular dichroism and crosslinking studies of the interaction between four neurotrophins and the extracellular domain of the low‐affinity neurotrophin receptor

Timm

Ross²,

Neet

1994

Protein Science

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“…Primary melanocytes, originating from the neural crest, and melanoma cell lines normally co-express p75NTR and TNF receptors but have little or no TrkA expression (43,44). Fig.…”

Section: Resultsmentioning

confidence: 99%

The p75 Neurotrophin Receptor (p75NTR) Alters Tumor Necrosis Factor-mediated NF-κB Activity under Physiological Conditions, but Direct p75NTR-mediated NF-κB Activation Requires Cell Stress

Bhakar¹,

Roux²,

Lachance³

et al. 1999

Journal of Biological Chemistry

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The p75 neurotrophin receptor (p75NTR) has been linked to activation of the NF-B transcriptional complex in oligodendrocytes, Schwann cells, and PCNA cells. In this report, tumor necrosis factor (TNF)-and neurotrophin-mediated NF (nuclear factor)-B activation were compared in several cell lines. All cell types showed TNF-mediated activation of NF-B, but direct neurotrophin-dependent activation of NF-B was never observed under normal growth conditions. In PCNA cells, a modest nerve growth factor (NGF)-dependent induction of NF-B was detected but only after cells were subjected to severe stress. Although NGF binding did not directly activate NF-B under normal conditions, NGF consistently altered TNF-dependent NF-B activation in each cell type examined, and extended exposure to NGF and TNF always increased NF-B activation over that achieved with TNF alone. The increase in NF-B activity mediated by NGF correlated with reduced levels of IB␣; NGF added alone had no effect on IB␣ levels, but when added with TNF, NGF treatment significantly reduced IB␣ levels. We propose that modulation of cytokine receptor signaling is a significant physiological function of the p75 neurotrophin receptor and that previous reports of direct NF-B activation through p75NTR reflect this modulatory activity.

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“…The receptor gene is present as a single copy (10,11), which is expressed as a single 3.8-kilobase (kb) mRNA. The cloned NGF receptor gene is expressed in heterologous cells as a 75-kDa glycoprotein, similar in size to the receptor found in rat pheochromocytoma PC12 cells (6,8), human melanoma cells (12)(13)(14), human neuroblastoma cells (8), and cultured sympathetic neurons (8,9). Cross species Southern blots indicate that the receptor gene is highly conserved during evolution (15).…”

mentioning

confidence: 93%

Developmentally regulated expression of the nerve growth factor receptor gene in the periphery and brain.

Buck¹,

Martínez²,

Black³

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

110

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Nerve growth factor (NGF) regulates development and maintenance of function of peripheral sympathetic and sensory neurons. A potential role for the trophic factor in brain has been detected only recently. The ability of a cell to respond to NGF is due, in part, to expression of specific receptors on the cell surface. To study tissue-specific expression of the NGF receptor gene, we have used sensitive cRNA probes for detection of NGF receptor mRNA. Our studies indicate that the receptor gene is selectively and specifically expressed in sympathetic (superior cervical) and sensory (dorsal root) ganglia in the periphery, and by the septum-basal forebrain centrally, in the neonatal rat in vivo. Moreover, examination of tissues from neonatal and adult rats reveals a marked reduction in steady-state NGF receptor mRNA levels in sensory ganglia. In contrast, a 2-to 4-fold increase was observed in the basal forebrain and in the sympathetic ganglia over the same time period. Our observations suggest that NGF receptor mRNA expression is developmentally regulated in specific areas of the nervous system in a differential fashion.Formation of the nervous system is dependent on a complex interaction of genetic and epigenetic factors and signals. Nerve growth factor (NGF) is required for neuronal survival in vivo and elicits diverse responses, including neurite elongation, neurotransmitter enzyme induction, and target-specific innervation in responsive neurons. The many actions of NGF on responsive neurons are initiated by interaction with a specific cell membrane-bound receptor. The NGF-receptor complex is internalized and transported from nerve terminals to the neuronal cell body, where critical cellular processes are mediated (1). Kinetic analyses have indicated that there are high-and low-affinity forms of the receptor molecule (2-4). Finally, 125I-labeled NGF binding in conjunction with cross-linking agents has allowed characterization of the ligand-receptor complex (5-9).We The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Purification and Amino Terminal Sequencing of Human Melanoma Nerve Growth Factor Receptor

Cited by 84 publications

References 27 publications

Circular dichroism and crosslinking studies of the interaction between four neurotrophins and the extracellular domain of the low‐affinity neurotrophin receptor

Circular dichroism and crosslinking studies of the interaction between four neurotrophins and the extracellular domain of the low‐affinity neurotrophin receptor

The p75 Neurotrophin Receptor (p75NTR) Alters Tumor Necrosis Factor-mediated NF-κB Activity under Physiological Conditions, but Direct p75NTR-mediated NF-κB Activation Requires Cell Stress

Developmentally regulated expression of the nerve growth factor receptor gene in the periphery and brain.

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